Confirmation of Vpr as a fibrinolytic enzyme present in extracellular proteins of Bacillus subtilis

Chang Won Kho; Sung Goo Park; Sayeon Cho; Do Hee Lee; Pyung Keun Myung; Byoung Chul Park

doi:10.1016/j.pep.2004.08.008

Confirmation of Vpr as a fibrinolytic enzyme present in extracellular proteins of Bacillus subtilis

Chang Won Kho, Sung Goo Park, Sayeon Cho, Do Hee Lee, Pyung Keun Myung, Byoung Chul Park

Research output: Contribution to journal › Article › peer-review

34 Scopus citations

Abstract

We have previously reported a proteomic approach to detect fibrinolytic enzymes from the secreted proteins of Bacillus subtilis 168 and identified two extracellular fibrinolytic enzymes of Bacillus, namely, Vpr and WprA. In this study, to confirm the fibrinolytic activity of Vpr, we cloned the vpr gene and expressed it in Escherichia coli, where it is predominantly localized to inclusion bodies. After affinity purification and desalting steps, the expressed Vpr is auto-processed to an active form. Interestingly, after the desalting step, several additional bands with fibrinolytic activity were detected in zymography gel along with a mature form (68 kDa) of Vpr. MALDI-TOF analyses of these bands revealed that Vpr could exist in multiple forms.

Original language	English
Pages (from-to)	1-7
Number of pages	7
Journal	Protein Expression and Purification
Volume	39
Issue number	1
DOIs	https://doi.org/10.1016/j.pep.2004.08.008
State	Published - Jan 2005
Externally published	Yes

Keywords

Fibrinolytic enzymes
Mass spectrometry
Serine protease
Vpr
Zymography

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10.1016/j.pep.2004.08.008

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title = "Confirmation of Vpr as a fibrinolytic enzyme present in extracellular proteins of Bacillus subtilis",

abstract = "We have previously reported a proteomic approach to detect fibrinolytic enzymes from the secreted proteins of Bacillus subtilis 168 and identified two extracellular fibrinolytic enzymes of Bacillus, namely, Vpr and WprA. In this study, to confirm the fibrinolytic activity of Vpr, we cloned the vpr gene and expressed it in Escherichia coli, where it is predominantly localized to inclusion bodies. After affinity purification and desalting steps, the expressed Vpr is auto-processed to an active form. Interestingly, after the desalting step, several additional bands with fibrinolytic activity were detected in zymography gel along with a mature form (68 kDa) of Vpr. MALDI-TOF analyses of these bands revealed that Vpr could exist in multiple forms.",

keywords = "Fibrinolytic enzymes, Mass spectrometry, Serine protease, Vpr, Zymography",

author = "Kho, {Chang Won} and Park, {Sung Goo} and Sayeon Cho and Lee, {Do Hee} and Myung, {Pyung Keun} and Park, {Byoung Chul}",

note = "Funding Information: This research was supported by a grant from MOCIE to Intelligence Bioinformatics and Application Center, KRIBB.",

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AU - Kho, Chang Won

AU - Park, Sung Goo

AU - Cho, Sayeon

AU - Lee, Do Hee

AU - Myung, Pyung Keun

AU - Park, Byoung Chul

N1 - Funding Information: This research was supported by a grant from MOCIE to Intelligence Bioinformatics and Application Center, KRIBB.

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AB - We have previously reported a proteomic approach to detect fibrinolytic enzymes from the secreted proteins of Bacillus subtilis 168 and identified two extracellular fibrinolytic enzymes of Bacillus, namely, Vpr and WprA. In this study, to confirm the fibrinolytic activity of Vpr, we cloned the vpr gene and expressed it in Escherichia coli, where it is predominantly localized to inclusion bodies. After affinity purification and desalting steps, the expressed Vpr is auto-processed to an active form. Interestingly, after the desalting step, several additional bands with fibrinolytic activity were detected in zymography gel along with a mature form (68 kDa) of Vpr. MALDI-TOF analyses of these bands revealed that Vpr could exist in multiple forms.

KW - Fibrinolytic enzymes

KW - Mass spectrometry

KW - Serine protease

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KW - Zymography

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Confirmation of Vpr as a fibrinolytic enzyme present in extracellular proteins of Bacillus subtilis

Abstract

Keywords

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